Genetic Identification and Symbiotic Efficiency of an Indigenous Sinorhizobium meliloti Field Population

Mihaela Bradić1*, Sanja Sikora1, Sulejman Redžepović1 and Zvonimir Štafa2

Department of Microbiology, Faculty of Agriculture, University of Zagreb, Svetošimunska 25, HR-10 000 Zagreb, Croatia

2Department of Field Crops Forage and Grassland Faculty of Agriculture, University of Zagreb, Svetošimunska 25, HR-10 000 Zagreb, Croatia

Article history:

Received: December 2, 2002
Accepted: January 24, 2003

Key words:

nitrogen fixation, alfalfa, Sinorhizobium meliloti, PCR-RFLP, rep-PCR, RAPD, symbiotic efficiency


Soil bacteria Sinorhizobium meliloti are of enormous agricultural value, because of their ability to fix atmospheric nitrogen in symbiosis with an important forage crop legume – alfalfa. The main aim of this study was (i) to isolate indigenous S. meliloti strains from different field sites in Croatia, (ii) to assess genetic diversity and genetic relationships amongst strains of natural populations and (iii) to provide information about nodulation and symbiotic efficiency of indigenous S. meliloti strains. The nine strains isolated from alfalfa nodules collected from different field sites and three reference strains were analysed. Genetic characterisation by PCR-RFLP of the 16S rDNA, rep-PCR and RAPD-PCR was applied to study the status of Sinorhizobium meliloti populations inhabiting nodules of alfalfa. The results of PCR-RFLP of the 16S rDNA revealed that all isolates belong to the S. meliloti species. Cluster analysis of rep-PCR and RAPD-PCR profiles showed significant differences among S. meliloti isolates. Both methods resulted in identical grouping of strains. Among indigenous strains two divergent groups could be determined. The biggest differences were detected among two reference strains and all field isolates. Greenhouse studies were performed for evaluation of symbiotic efficiency and compatibility of S. meliloti strains with two alfalfa cultivars. Quantitative expression of symbiotic efficiency was evaluated by measurement of nodule dry weight, content of proteins and total nitrogen in plants, dry matter and green mass yield of plants. All strains nodulated both alfalfa cultivars but with different efficiency. Significant differences in dry matter and green mass yield of alfalfa as well as protein content were determined depending on the strain used. The results indicate that three indigenous S. meliloti strains can be characterised as the most efficient of all strains used in this study. 

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